Viruses have been responsible for many epidemics and pandemics that have impacted human life globally. The COVID-19 pandemic highlighted both our vulnerability to viral outbreaks, as well as the mobilization of the scientific community to come together to combat the unprecedented threat to humanity. Cryo-electron microscopy (cryo-EM) played a central role in our understanding of SARS-CoV-2 during the pandemic and continues to inform about this evolving pathogen. Cryo-EM with its two popular imaging modalities, single particle analysis (SPA) and cryo-electron tomography (cryo-ET), has contributed immensely to understanding the structure of viruses and interactions that define their life cycles and pathogenicity. Here, we review how cryo-EM has informed our understanding of three distinct viruses, of which two - HIV-1 and SARS-CoV-2 infect humans, and the third, bacteriophages, infect bacteria. For HIV-1 and SARS-CoV-2 our focus is on the surface glycoproteins that are responsible for mediating host receptor binding, and host and cell membrane fusion, while for bacteriophages, we review their structure, capsid maturation, attachment to the bacterial cell surface and infection initiation mechanism.

The basic building block of the cerebral cortex, the pyramidal cell, has been shown to be characterized by a markedly different dendritic structure among layers, cortical areas, and species. Functionally, differences in the structure of their dendrites and axons are critical in determining how neurons integrate information. However, within the human cortex, these neurons have not been quantified in detail. In the present work, we performed intracellular injections of Lucifer Yellow and 3D reconstructed over 200 pyramidal neurons, including apical and basal dendritic and local axonal arbors and dendritic spines, from human occipital primary visual area and associative temporal cortex. We found that human pyramidal neurons from temporal cortex were larger, displayed more complex apical and basal structural organization, and had more spines compared to those in primary sensory cortex. Moreover, these human neocortical neurons displayed specific shared and distinct characteristics in comparison to previously published human hippocampal pyramidal neurons. Additionally, we identified distinct morphological features in human neurons that set them apart from mouse neurons. Lastly, we observed certain consistent organizational patterns shared across species. This study emphasizes the existing diversity within pyramidal cell structures across different cortical areas and species, suggesting substantial species-specific variations in their computational properties.

Forensic Facial Approximation (FFA) has evolved, with techniques advancing to refine the intercorrelation between the soft-tissue facial profile and the underlying skull. FFA has become essential for identifying unknown persons in South Africa, where the high number of migrant and illegal labourers and many unidentified remains make the identification process challenging. However, existing FFA methods are based on American or European standards, rendering them inapplicable in a South African context. We addressed this issue by conducting a study to create prediction models based on the relationships between facial morphology and known factors, such as population affinity, sex, and age, in white South African and French samples. We retrospectively collected 184 adult cone beam computed tomography (CBCT) scans representing 76 white South Africans (29 males and 47 females) and 108 French nationals (54 males and 54 females) to develop predictive statistical models using a projection onto latent structures regression algorithm (PLSR). On training and untrained datasets, the accuracy of the estimated soft-tissue shape of the ears, eyes, nose, and mouth was measured using metric deviations. The predictive models were optimized by integrating additional variables such as sex and age. Based on trained data, the prediction errors for the ears, eyes, nose, and mouth ranged between 1.6 mm and 4.1 mm for white South Africans; for the French group, they ranged between 1.9 mm and 4.2 mm. Prediction errors on non-trained data ranged between 1.6 mm and 4.3 mm for white South Africans, whereas prediction errors ranging between 1.8 mm and 4.3 mm were observed for the French. Ultimately, our study provided promising predictive models. Although the statistical models can be improved, the inherent variability among individuals restricts the accuracy of FFA. The predictive validity of the models was improved by including sex and age variables and considering population affinity. By integrating these factors, more customized and accurate predictive models can be developed, ultimately strengthening the effectiveness of forensic analysis in the South African region.

AMPA-type glutamate receptors (AMPAR) mediate excitatory cochlear transmission. However, the unique roles of AMPAR subunits are unresolved. Lack of subunit GluA3 (Gria3KO) in male mice reduced cochlear output by 8-weeks of age. Since Gria3 is X-linked and considering sex differences in hearing vulnerability, we hypothesized accelerated presbycusis in Gria3KO females. Here, auditory brainstem responses (ABR) were similar in 3-week-old female Gria3WT and Gria3KO mice. However, when raised in ambient sound, ABR thresholds were elevated and wave-1 amplitudes were diminished at 5-weeks and older in Gria3KO. In contrast, these metrics were similar between genotypes when raised in quiet. Paired synapses were similar in number, but lone ribbons and ribbonless synapses were increased in female Gria3KO mice in ambient sound compared to Gria3WT or to either genotype raised in quiet. Synaptic GluA4:GluA2 ratios increased relative to Gria3WT, particularly in ambient sound, suggesting an activity-dependent increase in calcium-permeable AMPARs in Gria3KO. Swollen afferent terminals were observed by 5-weeks only in Gria3KO females reared in ambient sound. We propose that lack of GluA3 induces sex-dependent vulnerability to AMPAR-mediated excitotoxicity.

New advanced technologies have recently been developed and preliminarily applied to surgery, including virtual reality (VR), augmented reality (AR) and mixed reality (MR). We retrospectively review all colorectal cases in which we used holographic 3D reconstruction from February 2020 to December 2022. This innovative approach was used to identify vascular anomalies, pinpoint tumor locations, evaluate infiltration into neighboring organs and devise surgical plans for both training and educating trainee assistants. We have also provided a state-of-the-art analysis, briefly highlighting what has been stated by the scientific literature to date. VR facilitates training and anatomical assessments, while AR enhances training and laparoscopic performance evaluations. MR, powered by HoloLens, enriches anatomic recognition, navigation, and visualization. Successful implementation was observed in 10 colorectal cancer cases, showcasing the effectiveness of MR in improving preoperative planning and its intraoperative application. This technology holds significant promise for advancing colorectal surgery by elevating safety and reliability standards.

Single-particle imaging using X-ray free-electron lasers (XFELs) is a promising technique for observing nanoscale biological samples under near-physiological conditions. However, as the sample\'s orientation in each diffraction pattern is unknown, advanced algorithms are required to reconstruct the 3D diffraction intensity volume and subsequently the sample\'s density model. While most approaches perform 3D reconstruction via determining the orientation of each diffraction pattern, a correlation-based approach utilizes the averaged spatial correlations of diffraction intensities over all patterns, making it well suited for processing experimental data with a poor signal-to-noise ratio of individual patterns. Here, a method is proposed to determine the 3D structure of a sample by analyzing the double, triple and quadruple spatial correlations in diffraction patterns. This ab initio method can reconstruct the basic shape of an irregular unsymmetric 3D sample without requiring any prior knowledge of the sample. The impact of background and noise on correlations is investigated and corrected to ensure the success of reconstruction under simulated experimental conditions. Additionally, the feasibility of using the correlation-based approach to process incomplete partial diffraction patterns is demonstrated. The proposed method is a variable addition to existing algorithms for 3D reconstruction and will further promote the development and adoption of XFEL single-particle imaging techniques.

The first land ecosystems were composed of organisms considered simple in nature, yet the morphological diversity of their flora was extraordinary. The biological significance of this diversity remains a mystery largely due to the absence of feasible study approaches. To study the functional biology of Early Devonian flora, we have reconstructed extinct plants from fossilised remains in silico. We explored the morphological diversity of sporangia in relation to their mechanical properties using finite element method. Our approach highlights the impact of sporangia morphology on spore dispersal and adaptation. We discovered previously unidentified innovations among early land plants, discussing how different species might have opted for different spore dispersal strategies. We present examples of convergent evolution for turgor pressure resistance, achieved by homogenisation of stress in spherical sporangia and by torquing force in Tortilicaulis-like specimens. In addition, we show a potential mechanism for stress-assisted sporangium rupture. Our study reveals the deceptive complexity of this seemingly simple group of organisms. We leveraged the quantitative nature of our approach and constructed a fitness landscape to understand the different ecological niches present in the Early Devonian Welsh Borderland flora. By connecting morphology to functional biology, these findings facilitate a deeper understanding of the diversity of early land plants and their place within their ecosystem.

Faced with the environmental challenges posed by climate change, architects are creating nature-based solutions for urban areas, such as transforming living trees into artificial architectural structures. In this study, we have analyzed stem pairs of five tree species conjoined for more than eight years by measuring the stem diameters below and above the resulting inosculation and by calculating the respective diameter ratio. Our statistical analyses reveal that Platanus × hispanica and Salix alba stems do not differ significantly in diameter below inosculation. However, in contrast to P. × hispanica, the diameters of the conjoined stems above inosculation differ significantly in S. alba. We provide a binary decision tree based on diameter comparisons above and below inosculation as a straightforward tool for identifying the likelihood of full inosculation with water exchange. Moreover, we have compared branch junctions and inosculations by means of anatomical analyses, micro-computed tomography, and 3D reconstructions showing similarities in the formation of common annual rings that increase the capacity for water exchange. Due to the highly irregular cell arrangement in the center of the inosculations, cells cannot be assigned clearly to either of the stems. In contrast, cells in the center of branch junctions can always be attributed to one of the branches.

Cochlear sound encoding depends on α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptors (AMPARs), but reliance on specific pore-forming subunits is unknown. With 5-week-old male C57BL/6J Gria3-knockout mice (i.e., subunit GluA3KO) we determined cochlear function, synapse ultrastructure, and AMPAR molecular anatomy at ribbon synapses between inner hair cells (IHCs) and spiral ganglion neurons. GluA3KO and wild-type (GluA3WT) mice reared in ambient sound pressure level (SPL) of 55-75 dB had similar auditory brainstem response (ABR) thresholds, wave-1 amplitudes, and latencies. Postsynaptic densities (PSDs), presynaptic ribbons, and synaptic vesicle sizes were all larger on the modiolar side of the IHCs from GluA3WT, but not GluA3KO, demonstrating GluA3 is required for modiolar-pillar synapse differentiation. Presynaptic ribbons juxtaposed with postsynaptic GluA2/4 subunits were similar in quantity, however, lone ribbons were more frequent in GluA3KO and GluA2-lacking synapses were observed only in GluA3KO. GluA2 and GluA4 immunofluorescence volumes were smaller on the pillar side than the modiolar side in GluA3KO, despite increased pillar-side PSD size. Overall, the fluorescent puncta volumes of GluA2 and GluA4 were smaller in GluA3KO than GluA3WT. However, GluA3KO contained less GluA2 and greater GluA4 immunofluorescence intensity relative to GluA3WT (threefold greater mean GluA4:GluA2 ratio). Thus, GluA3 is essential in development, as germline disruption of Gria3 caused anatomical synapse pathology before cochlear output became symptomatic by ABR. We propose the hearing loss in older male GluA3KO mice results from progressive synaptopathy evident in 5-week-old mice as decreased abundance of GluA2 subunits and an increase in GluA2-lacking, GluA4-monomeric Ca2+-permeable AMPARs.

The study aimed to describe the morphogenesis of the domestic cat uterine tube, from the primordial tubal segment of the paramesonephric duct (TsPD) to the mature organ. The specific aims included identifying differences in the rate and way of development of the layers in the wall of the infundibulum, the ampulla, and the isthmus. In the study we conducted light microscopic (LM) observations of the uterine tube wall, and analysis of 3D models, which were used for the first time to visualize the arrangement of the paramesonephric duct and highlight the pattern of the developing mucosal folds. The results revealed the two stages in the prenatal development of the domestic cat uterine tube. The first stage lasts from day 26-44 p.c. (post-conception) and includes the growth of the TsPD. It starts with formation of the TsPD\'s epithelial tubule in the mesenchyme of the anterior part of the mesonephros. The tubule, surrounded by mesenchyme, elongates posteriorly, and the TsPD can be divided into three sections according to their location and diameter, i.e., anterior, middle, and posterior. The first stage ends with the histodifferentiation of the mesenchyme into loose connective tissue and mucosa formation. The second stage lasts from approximately day 45-63 p.c. and comprises the formation of the proper layers in the uterine tube wall. The pace and pattern of layers\' development are specific to each section of the uterine tube, as tissues first differentiate in its anterior part and then continue posteriorly. The mucosal folds appear after day 44 p.c. in the infundibulum, and they become long and branched by the end of the prenatal period. Then, approximately day 48 p.c., the single mucosal folds appear in the ampulla, and at the end, i.e., about day 63 p.c., they become visible in the isthmus. The 3D models revealed the unique spiral-shaped arrangement of the mucosal folds in the ampulla. The tunica muscularis starts to develop ca day 48-51 p.c. in the anterior part of the uterine tube. On approximately day 55 p.c., the circular muscle layer can be distinguished in the ampulla, while in the isthmus, it develops just before birth. The postnatal development of the infundibulum includes the formation of the labyrinth-like system of mucosal folds, whereas in the ampulla and isthmus, branched mucosal folds and the second longitudinal muscle layer develop postnatally. At the end of the prenatal period, the differences in the wall thickness between the individual segments of the uterine tube reflect the proper proportions of the uterine tube wall in the mature female.